Reliable reference genes for expression analysis of proliferating and adipogenically differentiating human adipose stromal cells.
Cell Mol Biol Lett
; 24: 14, 2019.
Article
em En
| MEDLINE
| ID: mdl-30815013
ABSTRACT
BACKGROUND:
The proliferation and adipogenic differentiation of adipose stromal cells (ASCs) are complex processes comprising major phenotypical alterations driven by up- and downregulation of hundreds of genes. Quantitative RT-PCR can be employed to measure relative changes in the expression of a gene of interest. This approach requires constitutively expressed reference genes for normalization to counteract inter-sample variations due to differences in RNA quality and quantity. Thus, a careful validation of quantitative RT-PCR reference genes is needed to accurately measure fluctuations in the expression of genes. Here, we evaluated candidate reference genes applicable for quantitative RT-PCR analysis of gene expression during proliferation and adipogenesis of human ASCs with the immunophenotype DLK1+/CD34+/CD90+/CD105+/CD45-/CD31-.METHODS:
We evaluated the applicability of 10 candidate reference genes (GAPDH, TBP, RPS18, EF1A, TFRC, GUSB, PSMD5, CCNA2, LMNA and MRPL19) using NormFinder, geNorm and BestKeeper software.RESULTS:
The results indicate that EF1A and MRPL19 are the most reliable reference genes for quantitative RT-PCR analysis of proliferating ASCs. PSMD5 serves as the most reliable endogenous control in adipogenesis. CCNA2 and LMNA were among the least consistent genes.CONCLUSIONS:
Applying these findings for future gene expression analyses will help elucidate ASC biology.Palavras-chave
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Perfilação da Expressão Gênica
/
Gordura Abdominal
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Reação em Cadeia da Polimerase em Tempo Real
Limite:
Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article